Animals ; Autophagy ; Biomarkers, Tumor ; metabolism ; Caveolin 1 ; deficiency ; metabolism ; Drug Delivery Systems ; Humans ; Myofibroblasts ; pathology ; Neoplasms ; metabolism ; pathology ; Prognosis ; Stromal Cells ; metabolism ; Tumor Microenvironment

Objective To isolate and identify stem-like cells (CSCs) from human ACHN renal cell carcinoma cell line.Methods ACHN cancer stem cells CSCs were enriched by serum-free culture condition.Methyl thiazolyl tetrazolium (MTT) method was performed to draw the growth curve.Flow cytometry was performed to evaluate the expressions of two CSCs markers,CD133 and CD44.Real-time polymerase chain reaction (RT-PCR) was performed to evaluate the expression of stem-related genes (Oct3/4,Bmi,Nanog,and 3-catenin).The tumorigenicity in vivo was also examined.Results ACHN CSCs were formed by sphere-forming culturing.The data demonstrated that there were stronger capacities of proliferation in ACHN tumor spheres than that of ACHN cells.And the expressions of CD133 and CD44 were also significandy enhanced in tumor spheres.The data showed that four genes (Oct3/4,Bmi,Nanog,and β-catenin)were upregulated in sphere-forming cells.Sphere-forming cells had a higher tumorigenic potential in vivo than incubated cells.Conclusions Human ACHN renal cell carcinoma cell line sphere-forming cells can be enriched by serum-free culture condition and exhibits several characteristic cancer stem cells properties.

Objective Bone marrow mesenchymal stem cells ( BMSCs) , a kind of stem cells with multiple differentiation po-tentials, exist in the bone marrow and other organizations.This study aimed to investigate the repairing effect of the exogenous basic fi-broblast growth factor ( bFGF) against chronic obstructive pulmonary disease ( COPD) and its action mechanism, and to determine the expression of the bFGF gene in transfected rat BMSCs. Methods BMSCs were isolated, cultured and identified.The recombinant plasmid bFGF-pcDNA3.1 was constructed and sequenced.Liposome-mediated bFGF-pcDNA3.1 plasmid was transfected into the BM-SCs of the rat (bFGF-pcDNA3.1 transfection group), liposome-mediated pcDNA3.1 transfected into the BMSCs (pcDNA3.1 transfec-tion group) , and untransfected BMSCs used as the control.G418 screening was performed for 14 days.The gene and protein expres-sions of bFGF were determined by qRT-PCR and Western blot. Results The full-length sequence of the bFGF gene was consistent with that of the GenBank.The expression of the bFGF gene was significantly higher in the bFGF-pcDNA3.1 transfection group (7.028 ±0.568) than in the pcDNA3.1 transfection group (1.000 ±0.082) and the non-transfection control (1) (P<0.01), but with no statistically significant difference between the latter two groups (P>0.05).The expression of the bFGF protein was also re-markably higher in the bFGF-pcDNA3.1 transfection group (1.017 ±0.054) than in the pcDNA3.1 transfection group (0.217 ± 0.009) and the non-transfection control (0.165 ±0.013) (P<0.05), with no statistically significant difference between the latter two groups (P>0.05). Conclusion Mediated by the liposome reagent, the recombinant eukaryotic expression vector bFGF-pcD-NA3.1 can be transfected into rat BMSCs and expresses the bFGF gene and protein.

This article was aimed to study the effects of plumbagin to human hepatic stellate cells.Observations were made on the influence of proliferation inhibition rate,apoptosis,secretion of extracellular matrix function and matrix metalloproteinase (MMP) by plumbagin.HSC-LX2 and drug were co-incubated for 48 hours.Then,MTT assay was used in the detection of inhibition of cell proliferation.The flow cytometry was used in the detection of apoptosis.Immunohistochemical method was used to observe typeⅠ and Ⅲ collagen,MMP-1 and MMP-13 expression location and area.The results showed that the low,medium and high concentrations of plumbagin inhibited cell proliferation rate of HSC-LX2,induced apoptosis of cells,reduced the secretion of typeⅠ and Ⅲ collagen,and increased the secretion ability of MMP-1 and MMP-13.Effects mentioned above were dose-dependent with statistical difference (P < 0.05).Effects in the medium and high concentrations groups were stronger than colchicine group.It was concluded that plumbagin had the ability to inhibit cell proliferation rate of HSC-LX2,induce apoptosis,reduce the secretion of extracellular matrix,and increase the secretion ability of fibrin degradation enzyme.Therefore,it had intervention effect on the process of liver fibrosis.All effects mentioned above were dose-dependent.And effects in the medium and high concentrations groups were stronger than colchicine group.

OBJECTIVETo observe the efficacy and safety of Qizhi Jiangtang Capsule (QJC) in treating stage 3b diabetic kidney disease (DKD) patients with macroalbuminuria.

METHODSPatients who conformed to the diagnostic criteria of stage 3b DKD were randomly assigned to two groups according to random digital table, the experiment group and the control group, 84 in each group. All patients received a two-week elution period, and then were treated with basic Western therapy. Patients in the experiment group took QJC, 5 pills per time, 3 times a day, while those in the control group took Valsartan Capsule 160 mg each time, once daily. The observation period of follow-ups was limited within 6 months, and the time points were set as the baseline, 1st month, 3rd month, and 6th month. Systolic blood pressure (SBP), diastolic blood pressure (DBS), 24 h urine protein quantitative (24 h UPQ), plasma albumin (ALB), and serum creatinine (SCr) were detected and recorded, and estimated glomerular filtration rate (eGFR) was calculated. The occurrence of hypoglycemic reaction, coagulation disorder, gastrointestinal tract reaction, allergy, hyperkalemia, doubling of creatinine, and overall adverse events were observed and recorded at same time.

RESULTSFinally 81 patients in the experiment group and 80 patients in the control group were effectively included. Compared with the baseline level, SBP and DBS obviously decreased in the control group at month 1 of treatment (P < 0.05), and more significantly decreased at month 6 of treatment (P < 0.01). SBP at month 1, 3, and 6 of follow-ups; DBS at month 6 of follow-ups was lower in the control group than in the experiment group (P < 0.05). At month 1, 3, and 6 of follow-ups, 24 h UPQ of the experiment group was significantly lower than the baseline level (P < 0.01). It was also significantly lower than the level of the control group at the same time point (P < 0.05). There was no significant difference in 24 h UPQ at month 1, 3, and 6 of follow-ups between the control group and the baseline level (P > 0.05). ALB of the experiment group showed an increasing trend. It was significantly higher than the baseline level at month 6 (P < 0.05), which was also higher than that of the control group at same period (P < 0.05). There was no significant difference in the ALB level in the control group (P > 0.05). SCr of two groups showed an increasing trend. SCr of the experiment group was significantly higher at month 1, 3, and 6 follow-ups than the baseline level (P < 0.05). But the increment of SCr was higher in the control group than in the experimental group, and obviously higher than the baseline levels (P < 0.05). eGFR of both groups showed a decreasing trend. The decrement was higher in the control group than in the experimental group (P < 0.05). The proportion of progression of renal functions at month 1, 3, and 6 of follow-ups in the experimental group was 0.0% (0 case), 9.55% (8 cases), and 21.4% (18 cases), while they were 8.3% (7 cases), 21.4% (18 cases), and 40.5% (34 cases) in the control group. There was no statistical difference in the proportion of progression of renal functions between the two groups at month 3 and 6 of follow-ups (P < 0.05). There was no statistical difference in the incidence of adverse reactions between two groups (P > 0.05).

CONCLUSIONQJC could effectively reduce urinary protein of patients with stage 3b DKD, and delay the progression of renal functions.

Adult ; Albumins ; analysis ; Albuminuria ; drug therapy ; Blood Pressure ; drug effects ; Creatinine ; blood ; Diabetic Nephropathies ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glomerular Filtration Rate ; Humans ; Male ; Middle Aged ; Tetrazoles ; therapeutic use ; Treatment Outcome ; Valine ; analogs & derivatives ; therapeutic use ; Valsartan

Objective To investigate clinical manifestations and clinical diagnosis and treatment and pathological and immunohistochemical features in gastrointestinal stromal tumors. Methods The clinical data of fifty-two cases with gastrointestinal stromal tumors were collected, whose clinical diagnosis and treat-ment and pathological features were retrospectively analyzed from January 1995 to December 2007. Results All patients received operation therapy, only forty-five cases with complete surgical resection. The immu-nohistochemical staining showed that the cases with CD117 positive accounted for 100% (52/52) and CD34 positive accounted for 88.5% (46/52). Conclusions Surgery was necessary for all patients, especially complete surgical resection. Gastrointestinal stromal tumors were poor in preoperative diagnosis, which diag-nosis was based on the immunohistochemical staining of the tumor tissue. CD117 and CD34 tumor markers may help to diagnose gastrointestinal stromal tumors.

Objective To investigate the thyroid ultrastructural pathological changes of diabetes mellitus (DM) rats as well as the intervention effects of insulin and aminoguanidine. Methods Totally 87 rats were treated with streptozotocin to establish DM animal models and divided into DM group(n=27),insulin intervention group(n=32) and aminoguanidine intervention group(n=28),25 rats were taken as normal controls. Twelve and 20 weeks after the animal model establishment, animals were sacrificed, thyroid tissue was taken and ultrastructure was observed. Results In the thyroid of DM rats, follicular epithelial cells present as applanate shape, microvilli were depleted, rough endoplasmic reticulum dilated to irregular vesicular. None pinocytotic vacuole and casual primary or secondary lysosome were seen. Follicular cavity was dilated, colloid in the cavity had higher electronic-density. Interstitial edema, capillary base lamian was thickened at different stage. Proteo-substance deposition with granulo-shape, cloud shape or homogeneity appeared. The number of thyroid parafollicular cells increased. But endocrine granule in parafollicular cells was few. When compared with DM group, the thyroid tissue injury of insulin intervention group and aminoguanidine intervention group were lessened to different degree. Conclusion The hypofunctional thyroid follicular cells, large quantity of proteo-substance deposition in the interstitium and increased parafollicular cells of DM rats may be related with hyperglycemia toxicity. Insulin and aminoguanidine treatment have some protection effects.

BACKGROUND:MicroRNAs (miRNA) through regulating specific target gene mRNA expression play important roles in different processes of diseases. OBJECTIVE:To study the interaction of miRNA-21 with its target gene TLR4 in Hela cels. METHODS:The candidate target gene of miRNA-21 was determined according to miRNA analysis databases. The constructed recombinant adenovirus vector carrying pri-miRNA-21 gene was used, which could package and amplify viruses to transfect Hela cels. Then, the expression of fluorescent proteins was detected. Forty-eight hours after transfection of miRNA-21 or control, extracted proteins were used for detection of TLR4 protein using western blot assay. RESULTS AND CONCLUSION:Recombinant adenoviruses pAd/pri-miRNA-21 and pAd/neg at 100 MOI could successfuly infect Hela cels. Bioinformatic analysis suggested several possible binding sites between miRNA-21 and TLR4. The experimental results showed that miRNA-21 down-regulated TLR4 at protein levels, indicating that miRNA-21 can interfere with the expression of TLR4 target gene.

Objective To investigate the changes of red blood cells (RBC) and blood lipid in Han high altitude immigration dur‐ing the process of altitude acclimatization and their correlation .Methods Forty male Han nationality teachers entering Xizang Ali area and working for 1 year were selected .The blood samples were collected before entering plateau and within 3 d after returning to plain .The blood routine and the blood lipid level were detected .Results In the early return to plain ,the RBC ,TG ,CHO and LDL‐C levels in the research subjects were increased ,in which RBC and TG were significantly increased (P< 0 .01) ,HDL‐C was significantly decreased .After returning to plain ,RBC was positively correlated with TG and CHO (r=0 .46 ,P<0 .01 and r=0 .36 , P<0 .05) .Conclusion High altitude hypoxia environment is the primary cause leading to the change of RBC and blood lipid inde‐xes in the plain population after entering the plateau environment .

Background Epidemiological investigation showed that 15％-29％ of patients with cataract have preexisting astigmatism of ＞ 1.50 D.So to control astigmatism is very important to the improvement of visual function after cataract surgery.The implantation of Toric intraocular lens (IOL) is a new option for the correction of preexisting astigmatism during cataract surgery,now.Short-term clinical studies of cataract patients with AcrySof Toric IOL implantation have revealed a good stability.However,the evaluation of long-term clinical result is seldom.Objective This study was to evaluate the long-term clinical results of Toric intraocular lens(IOL) implantation.Methods A serial case-observational study was designed.One hundred and twenty eyes of 78 cataract patients were included in this study.Phacoemulsification combined with AcrySof Toric IOL implantation was performed and the patients received a 2-year follow-up.Uncorrected distance visual acuity (UCDVA),best corrected distance visual acuity(BCDVA),residual cylinder,IOL rotation,vector analysis and accuracy of astigmatic correction were clinically evaluated in 1 day,1 month,3 months,6 months,1 year and 2 years,respectively.Comparison of these results of different follow-up periods were made.Results Sixty-seven patients (101 eyes)finished the follow-up and 19patients(19 eyes)lost visit due to other diseases affected.At 2 years visit after AcrySof SN60TT implantation,UCDVA,BCDVA,residual cylinder,absolute value of IOL rotation degree,vector magnitude of surgically induced astigmatism (SIA) was 0.16 (0.20),0 (0.1),0.75 (0.5) D,(2.9± 1.8) °,(1.2 ± 0.6) D,and the correction index (CI)was 0.90±0.41.A positive correlation was found between SIA and TIA(r=0.74,P =0.000).Compared to 1 month,3,6 months and 1 year,there was a mild tendency of decrease in accuracy of astigmatism correction and CI.Residual cylinder and degree of IOL rotation at 2 years after surgery were also slightly higher.However these changes were not statistically significant (P＞0.05).Conclusions After 2 years of follow-up,patients with AcrySof Toric implantation remain good visual acuity.AcrySof Toric IOL presents excellent long-term rotational stability and accuracy of astigmatism correction.